Rapid detection method of nitrite in blood and urine with high sensitivity

A technology of nitrite and detection method is applied in the direction of material analysis by observing the impact on chemical indicators, and analysis by causing chemical reactions to occur in materials, etc., which can solve problems such as interference and difficulty in detecting nitrite, etc., To achieve the effect of mild operating conditions, short extraction time and high extraction rate

Inactive Publication Date: 2015-03-25
云南健牛环境监测有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Griess detection kit is usually used for detection. Due to the limitation of detection sensitivity and matrix interference, it is usually difficult to detect nitrite, and its sensitivity can only reach 1 μg / mg.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The invention is used to detect the content of nitrite in human urine.

[0023] 1. Production of nitrite standard color scale: take NO with a concentration of 5.0 μg / mL 2 - Put the standard solution 0.005, 0.01, 0.05, 0.10, 0.30, 0.70, 1.0mL in a graduated centrifuge tube, dilute to 5mL with distilled water, add 250μL of GriessA reagent, mix well; add 250μL of GriessB reagent after 1min, mix well, let stand for 5min, Add non-ionic surfactant AE0-9 0.1mL, ammonium sulfate 0.2g, vortex mix for 1min, centrifuge at 5000r / min for 10min and then separate the phases, and a red ring color solution from light to dark appears on the upper layer of the solution;

[0024] 2. Sample processing and measurement results

[0025] Take 10mL of urine into a 10mL centrifuge tube, add 1.0g of activated carbon and 0.2g of ammonium sulfate, mix well, centrifuge at 4000r / min for 10min, filter through filter paper to remove activated carbon and protein precipitates. Take 5.0 mL of dec...

Embodiment 2

[0027] The invention is used to detect the content of nitrite in human blood.

[0028] 1. make the standard color scale of nitrite by embodiment 1, nonionic surfactant is AEO-7, n-octanol 0.05mL replaces ammonium sulfate.

[0029] 2. Sample processing and measurement results

[0030] Take 5 mL of fresh blood and centrifuge at 6000r / min for 12min. Take the supernatant in a centrifuge tube, add 0.2mL of 0.1mol / L sodium hydroxide solution and 0.2g of zinc sulfate, mix well, add 1mL of acetonitrile, centrifuge at 4000r / min for 10min to separate, take 2mL of the supernatant into a 10mL centrifuge tube Dilute to 5 mL with distilled water, add 250 μL of GriessA reagent, mix well, add 250 μL of GriesssB reagent after 1 min, mix well, add non-ionic surfactant AEO-7 0.1 mL, n-octanol 0.05 mL, vortex mix for 1 min, 5000r / After centrifuging for 15 min, the phases were separated, and the color circle of the surfactant in the upper layer was compared with the standard color scale circle ...

Embodiment 3

[0032] The invention is used to detect the content of nitrite in human blood.

[0033] 1. Make the standard color scale of nitrite according to embodiment 1, the nonionic surfactant is AEO-3 organic alcohol and is n-amyl alcohol 0.1mL.

[0034] 2. Sample processing and measurement results

[0035]Take 5mL of fresh blood in a centrifuge tube, centrifuge at 6000r / min for 8min, take the supernatant in a centrifuge tube, mix 0.2mL of 0.1mol / L sodium hydroxide solution and 0.2g of zinc sulfate, add acetonitrile 1mL, 4000r / min Centrifuge for 10 min to separate, take 2 mL of the supernatant and dilute to 5 mL with distilled water in a 10 mL centrifuge tube, add 250 μL of GriessA reagent, and mix well. After 1 min, add 250 μL of GriessB reagent, mix well, and after standing for 5 min, add nonionic surfactant AEO-3 0.1 mL, n-pentanol 0.1 mL, vortex mix for 1 min, centrifuge at 5000 r / min for 10 min, and separate phases, the upper layer of surfactant Compared with the standard color c...

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Abstract

The invention discloses a rapid detection method of nitrite in blood and urine with high sensitivity. Azo compound is formed through diazotization coupling reaction by nitrite and a Griess agent. A nonionic surfactant including fatty alcohol polyoxyethylene ether AEO series is used in cloud point extraction. The method has advantages such as mild operation condition, low cloud point temperature, short phase-separation time, high extraction efficiency and the like. The detection limit of nitrite in the system can reach 0.005 micrograms / mL. According to the invention, the method with high sensitivity is simple and fast and can be used on site.

Description

technical field [0001] The invention belongs to the technical field of in vivo drug detection, in particular to a highly sensitive and rapid detection method for nitrite in blood and urine. Background technique [0002] Nitric oxide (NO) is a gas with unpaired electrons. It is chemically unstable and has a short half-life of only a few seconds. It is easy to form nitrite and nitrate. NO is the strongest vasodilation factor and contraction factor known so far, and it can participate in many physiological or pathological processes collectively as a mediator, messenger, transmitter or cell function regulator. NO is an important molecule that regulates the normal physiological activities of the myocardium. It has the functions of dilating blood vessels, inhibiting platelet aggregation and adhesion, anti-inflammation, and inhibiting cell apoptosis and proliferation. Humans have 60 trillion cells, and nitric oxide acts as a conduction factor between cells in the human body...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 杨亚玲王军赵娇宁金艳
Owner 云南健牛环境监测有限公司
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